|Year : 2018 | Volume
| Issue : 1 | Page : 42-46
Effect of Ferula assa foetida oleo-gum-resin on gastric ulcer in indomethacin-ulcerated rats
Seyyed Majid Bagheri1, Maryam Yadegari2, Fatemeh Zare-Mohazabiye1, Hanieh Momeni-Asl1, Aghdas Mirjalili3, Morteza Anvari3, Malihe Behpour1
1 Department of Physiology, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
2 Neurobiomedical Research Center, Shahid Sadoughi University of Medical Sciences; Department of Anatomy, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
3 Department of Anatomy, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
|Date of Submission||26-Aug-2017|
|Date of Acceptance||23-Oct-2017|
|Date of Web Publication||25-May-2018|
Seyyed Majid Bagheri
Department of Physiology, Shahid Sadoughi University of Medical Sciences, Prof. Hesabi Bulvd, Shohadaye Gomnam Bulvd, 8915173149 Yazd
Source of Support: None, Conflict of Interest: None
Background: Asafoetida is used as an antiulcer agent in Iran and Afghanistan. This study was undertaken to determine the healing of ulcers induced by indomethacin (IND) due to the protective role of asafoetida on ulcers in rats. Materials and Methods: Rats were divided into four groups with five rats in each group. One group only received IND as negative control group and other group received normal saline and considered as control group, respectively. Two groups were pretreated by asafoetida (25 and 50 mg/kg) by oral gavage for 4 days, and after this time, rats were kept fasted for 12 h and received IND at 48 mg/kg orally. Four hour later, animals were killed and their stomach and liver were fixed in formalin (10%) and sections of 5 μm in diameter were prepared. Histological and morphological characteristics of stomach and liver were assessed using hematoxylin and eosin (H and E) staining. Results: Treatment groups with asafoetida (25 and 50 mg/kg) showed significant (P < 0.05) reduction in microscopic and macroscopic ulcer index as compared to the IND control group. Asafoetida promotes ulcer protection as ascertained histologically by comparatively decrease in ulcer areas, reduction or absence of edema and leukocytes infiltration of submucosal layer compared to IND control group. The results also indicated that asafoetida has a protective effect on liver tissues. In general, ameliorative effects of asafoetida in animals treated at 50 mg/kg were better than dose of 25 mg/kg, respectively. Conclusions: Data indicates that asafoetida has anti-ulcer and hepatoprotective activity that these beneficial effects may be or mainly related to anticholinergic antihistaminergic and antioxidant effects.
Keywords: Asafoetida, hepatotoxicity, indomethacin, ulcer peptic
|How to cite this article:|
Bagheri SM, Yadegari M, Zare-Mohazabiye F, Momeni-Asl H, Mirjalili A, Anvari M, Behpour M. Effect of Ferula assa foetida oleo-gum-resin on gastric ulcer in indomethacin-ulcerated rats. J Curr Res Sci Med 2018;4:42-6
|How to cite this URL:|
Bagheri SM, Yadegari M, Zare-Mohazabiye F, Momeni-Asl H, Mirjalili A, Anvari M, Behpour M. Effect of Ferula assa foetida oleo-gum-resin on gastric ulcer in indomethacin-ulcerated rats. J Curr Res Sci Med [serial online] 2018 [cited 2019 Sep 18];4:42-6. Available from: http://www.jcrsmed.org/text.asp?2018/4/1/42/233196
| Introduction|| |
Peptic ulcer is a major health hazard that influenced by the loss of the balance between various aggressive and defensive factors such as acid-pepsin secretion, mucosal barrier, cellular regeneration, and gastric mucosa. It is demonstrated that increase in oxidative stress is linked to the aggressive factor-induced gastric mucosal damage. Indomethacin (IND) is one the nonsteroidal anti-inflammatory drugs (NSAIDs) with anti-inflammatory, antipyretic, and pain-relieving properties, which is known to produce erosions, ulcerative lesions and petechial bleeding in the mucosa of stomach as serious side effects. Furthermore, the development of the gastric mucosal lesions induced by IND is mainly mediated through generation of oxygen free radicals  and depletion of endogenous prostaglandins through inhibition of the enzyme cyclooxygenase. In the recent years, tendency towards herbal medicine has been increased and people have recognized that use of many cultivated wild plants and its products have less toxic effects than synthetic drugs and are a good source for novel therapeutic agents. Plants of the genus Ferula belongs to the family of Apiaceae include about 130 species that distributed throughout central Asia and Mediterranean area.Ferula assa-foetida L. is one of the species that wildly grows in central area of Iran and important part of this plant and several other species of Ferula is an oleo-gum-resin asafoetida yielded from incisions in the stem and/or roots of these plants. In different traditional medicines, asafoetida is introduced as a valuable remedy for gastrointestinal disorders. In India, asafoetida is used for flatulence and treatment of stomach ache. American people use it orally as an anti-helminthic, and it is claimed to be a powerful antispasmodic. The hot water extract of F. assa foetida oleo-gum-resin is used for the treatment of hysteria and whooping cough in Afghanistan as traditional medicine. Recent pharmacological and biological studies have also shown several pharmacological activities such as antioxidant, antileishmanial, cancer chemopreventive, anticonvulsant  antidiabetic, antispasmodic, hypotensive, and antinociceptive. According to some studies, asafoetida may also have hepatoprotective effects against toxic agents. Researchers showed that a mixture of the methanol-insoluble fraction of asafoetida, fresh garlic, curcumin, shown antihepatotoxic activity versus aflatoxin B1-induced hepatotoxicity. Dandagi et al. also ]foetida, Momordica charantia Linn, and Nardostachys jatamansi have physicochemical and hepatoprotective activity. Keeping these folkloric claims and reports in view, the present study is attempted to evaluate the possible antiulcer and hepatoprotective effect of asafoetida in IND induced ulcerated rats.
| Materials and Methods|| |
Male albino rats (250–300 g) with 6–8 weeks old, bred in animal house of Medical School of Shahid Sadoughi University of Medical Sciences were selected. Animals were housed at controlled temperature (22 ± 2°C) with a 12 h-light/dark cycle and with standard lab chow and tap water ad libitum. The experiments reported in this study were carried out in accordance with ethical guidelines of Shahid Sadoughi University of Medical Sciences for the investigation of experimental animals.
Preparation of plant oleo-gum-resin
F. assa-foetida oleo-gum-resin was collected from Tabas region (Yazd province, Iran) during the summer and the plant species was botanically identified by Dr. Abbas Zarezadeh in Yazd Agricultural Research Center. The dried powder of asafoetida was soaked in distilled water overnight at room temperature, and the yielded suspension was used orally. Concentrations and dosages of the suspension were expressed as crude amount of the dried oleo-gum-resin used in preparing the stock solution.
To evaluate the protective effect of asafoetida, gastric ulcers were induced by a single oral dose of 48 mg/kg IND. The animals were randomly classified into four groups (5 rats per each): The control group received normal saline orally in a volume of 5 ml/kg. IND group in which gastric ulceration was induced by i.p. injection of a single dose of 48 mg/kg IND. IND + asafoetida 25 group in which animals pretreated with 25 mg/kg asafoetida orally, 4 days before IND administration. IND + asafoetida 50 in which animals pretreated with 50 mg/kg orally 4 days before IND administration. Rats were sacrificed in deep ether anesthesia 4 h after the oral administration of IND, and their stomachs and livers were immediately removed for quantifying the lesions and histopathological analysis.
Assessment of gastric mucosal lesions
Long lesions were counted and measured along their greater length, and petechial lesions were also counted. Each five petechial lesions were taken as 1 mm of ulcer. The sum of the total length long ulcers and petechial lesions in each group of rats was divided by its number to calculate the ulcer index (mm). The macroscopic curative ratio was determined by the following formula:
Macroscopic curative ratio (% protection) = (control ulcer index)−(test ulcer index)/control ulcer index ×100
Then, microscopic ulcer index, curative ratio (% protection) and morphological characteristics of gastric tissues were evaluated, and a mean index was calculated. A mean microscopic ulcer index and curative ratio were calculated by the formula
- Normal tissue = 0
- Local damage to gastric pits cells = 1
- Local damage to gastric glands = 2
- Deep damage to gastric glands = 3.
Microscopic ulcer index = (number of lesion 1) + (number of lesion 2) ×2+ (number of lesion3) ×3
Microscopic % Protection= (control ulcer index)-(test ulcer index)/control ulcer index ×100
Livers and stomach were quickly removed and fixed in 10% formalin, dehydrated in gradual ethanol (50%–100%), cleared in xylene, and embedded in paraffin. Sections (4–5 μm thick) were prepared and then stained with hematoxylin and eosin dye for photomicroscopic observations of the liver histologic architecture of the control and treated rats.
All data are expressed as the mean ± standard error of the means. GraphPad Prism 5 (GraphPad, San Diego, CA, USA) was used for data analysis. Statistically significant differences were determined using one-way ANOVA with the Tukey Kramer posttest for multiple comparisons. The values of P < 0.05 were regarded as statistically significant.
| Results|| |
Effect of extract on microscopic and macroscopic ulcer index and curative ratio
The results indicated that asafoetida significantly decreased the microscopic and macroscopic ulcer index (P < 0.05). Asafoetida also exerted microscopic and macroscopic curative ratios compare to IND group. Treatment with the asafoetida revealed a statistically significant improvement in the ulcer healing process in a dose-dependent manner [Table 1].
|Table 1: Effect of asafoetida 25 and 50 mg/kg pretreatment on ulcer index and % protection in indomethacin-induced gastric ulcer in rats|
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Effect of asafoetida on stomach histopathology
Histological observation of the gastric mucosa of control showed normal mucosa and muscularis mucosa structure [Figure 1]a. The stomachs of the IND treated rats, showed deep damage to gastric glands and mucosal hemorrhage and severe discontinuity and disruption in the lining epithelium. There is extensive edema of the submucosa and leukocytes infiltration in submucosal layer. Venule expansion in lamina propria also was seen, but muscularis mucosa was undamaged [Figure 1]b. Pretreated animals with the asafoetida on dose of 25 mg/kg showed mild discontinuity in the lining epithelium and submucosal edema. Venule expansion in lamina propria and submucosal layer and local damage to gastric glands was seen [Figure 1]c. Treatment with the asafoetida (50 mg/kg) resulted in the maintenance of glandular organization and cellular architecture. Asafoetida on a dose of 50 mg/kg, had great effect with no ulcer formation, thick muscularis mucosa. Less disruption to gastric mucosa layer with leukocyte infiltration was seen but there was no edema in the submucosa [Figure 1]d. The results indicated that asafoetida showed to exert the cytoprotective effects in a dose-dependent manner.
|Figure 1: Histology evaluation of gastric mucosal lesions in different groups H and E ×100. (a) Control group showed normal mucosa and muscularis mucosa structure. The lining epithelium was intact (a) There is severe disruption to the surface epithelium and edema of the submucosa layer with leukocyte infiltration in stomachs of the indomethacin-treated rats. (b) In stomach of rats treated with asafoetida (25 mg/kg), there is mild disruption to the surface epithelium mucosa, but there was edema and leukocytes infiltration of the submucosal layer. (c) In stomach of rats treated with asafoetida (50 mg/kg), reduced submucosal edema. There is no disruption to the surface epithelium, but there is less leukocytes infiltration of the submucosal layer. (d) Leukocytes infiltration and edema in the submucosal layer and muscularis mucosa were shown by arrowhead, arrow and #respectively|
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Effect of extract on liver histopathology
Histological examinations of liver sections under light microscope revealed that there were several changes in the liver treated with all doses of asafoetida as compared with the IND control group. In the normal liver tissue section (control group) showed normal hepatic architecture and hepatocytes with prominent nucleus. Normal cords of hepatocytes with central vein and portal tracts with no sign of inflammation or necrosis was observed [Figure 2]a. Whereas the liver tissue section in IND treated group showed distortion in the arrangement of cells around the central vein, inflammations around central vein, and leukocyte infiltration. Dilated hepatic sinusoids and vessels expansion in portal area were observed. Hypertrophied kupffer cells were prominent and hepatocytes were larger compared with control group [Figure 2]b. Asafoetida (25 and 50 mg/kg) treated animals brought back the cellular arrangement around the central vein and reduced hepatic sinusoids and vessels expansion in portal area and sinusoids. Asafoetida helped to bring the blood vessels to normal condition in a dose-dependent manner [Figure 2]c and [Figure 2]d. No histopathological changes visible in the sections of asafoetida (50 mg/kg) compared with control group.
|Figure 2: Histopathological changes in liver of control and experimental rats. H and E ×400. Control group: Control liver showing the central vein with radiating cords of hepatocytes (a) in indomethacin-treated rats disarrangement and degeneration of hepatocytes were observed. Dilated hepatic sinusoids (arrow) and leukocytes infiltrations (arrowhead) in these parts were noticed (b) in treated rat with the asafoetida (25 mg/kg), liver tissue showed normal radiating cords of hepatocytes, and there were no changes in central vein and portal areas. Hepatic sinusoids were normal, but leukocytes infiltration in these areas were seen (c) in rats treated with (50 mg/kg) asafoetida, hepatocytes structure and arrangement and hepatic sinusoids was normal. Mild dilated hepatic sinusoids in these areas were observed (d)|
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| Discussion|| |
The findings of present study demonstrated that asafoetida significantly decreased macroscopic and microscopic ulcer index and increased curative ratio or percent protection compare to IND control group and these effects were dose-dependent manner. The results on histological investigation on the gastric tissue of rats revealed that the pretreatment with asafoetida suspension significantly inhibited the IND various pathological indices. Among the different models of experimentally induced gastrointestinal lesions, the NSAID models is frequently used. The mechanisms suggested for the gastric damage caused by NSAIDs are inhibition of prostaglandin synthesis and inhibition of epithelial cell proliferation in the ulcer margin. Ulcer therapy has progressed from vagotomy to anticholinergic drugs, histamine H2 receptor antagonists, antacids and to proton pump inhibitors. Acetylcholine is one of the neurotransmitter that caused increasing secretion of acid and induced contraction through M3 subtype of muscarinic receptor in the gastrointestinal smooth muscle. It can be concluded that anticholinergic drugs have been shown to inhibit acid secretion and slow gastric motility and perhaps this could be the one of the mechanism(s) by which the suspension of asafoetida offers its ulcer protective effect. The inhibitory effect (antagonism effect) of asafoetida on muscarinic receptors was previously investigated and researchers showed that high concentration of asafoetida has noncompetitive antagonistic effect on muscarinic receptors. Previously, researchers also reported a potent antioxidant activity of asafoetida. It is well established that antioxidants play an important role in preventing gastric mucosal damage by strong cell defense mechanisms  are likely to stimulate the endogenous synthesis of prostaglandins, or by a protective role as a membrane stabilizing agent. In addition, one study showed that IND (a cyclooxygenase inhibitor) remarkably inhibited the relaxant effect of Ferula asafoetida gum which suggested that cyclooxygenase metabolites (prostaglandins such as PGE2, PGD2, and PGI2) might be involved in this relaxation. Histamine is one another of gastrointestinal system factors that its secretory effects apply through H1 receptor activation. The histamine receptor on parietal cells is the H2 type, and blocking the binding of histamine to this receptor is a widely used method for suppressing gastric acid secretion. Previous study showed that asafoetida has relaxant effects on isolated guinea-pig ileum preparations precontracted by histamine and suggested that asafoetida could block histamine receptors. In this study, we also investigated hepatoprotective effect of asafoetida on histological and morphological property of liver. It is demonstrated that IND caused degenerative changes histologically and cell necrosis in liver's cells and decrease of hepatic microsomal cytochrome P-450 and prostaglandin. Our histopathological results showed that in pretreated rat with asafoetida pathological changes such as leukocytes infiltration were not seen. Asafoetida suspension appears to possess significant cytoprotective activity as prior treatment of the suspension significantly inhibited the formation of liver damage induced by IND -in rats. These results support the hepatoprotective property of asafoetida could be related to a cytoprotective and it could also be through partly antioxidant and/or prostaglandin-dependent mechanism(s).
| Conclusions|| |
It appears that asafoetida possesses hepatoprotective and gastric antiulcer activity in rats. Further studies can also be conducted to determine the active constituent of asafoetida that gives antiulcer and hepatoprotective property to this oleo-gum-resin.
The authors thank all people who have assisted the experimental procedure and to the research deputy of Yazd Shahid Sadoughi Medical University, as the sponsor of this research.
Financial support and sponsorship
This research was supported by the foundation of Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran.
Conflicts of interest
There are no conflicts of interest.
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